Novel dammarane sapogenins, their use as anti-cancer agents, and a process for producing same

ABSTRACT

This invention relates to a group of novel sapogenins, their use in anti-cancer applications, and to a process for their production. More particularly, this invention pertains to a novel group of dammarane sapogenins, PAM-120, PBM-110 and PBM-100 (the dammarance sapogenine structure is specifically clean of any sugar moieties (glycons) at any position and hydroxyl at C-20) and PAN-20 and PAN-30 (the dammarance sapogenin structure has sugar moieties but is free of hydroxyl at C-20), obtained by chemical cleavage of dammarane saponins. The invention also includes a novel application of the said sapogenins for anti-cancer treatment by using them separately or together, and/or jointly with other drugs, as well as to the process of producing these novel sapogenins. Said novel dammarane sapogenins show surprising anti-cancer effect when applied, particularly against multi-drug resistant cancers.

[0001] This is a continuation-in-part of application Ser. No.09/910,887, filed Jul. 24, 2001.

FIELD OF THE INVENTION

[0002] This invention relates to novel dammarane sapogenins, their usein anti-cancer applications, and a process of producing the dammaranesapogenins. More particularly, the invention pertains to a novel groupof dammarane sapogenins obtained by chemical cleavage of dammaranesaponins extracted from panax ginseng, panax quinguefol, panaxnotoginseng and other species in the ginseng family, and a novelpreparation of anticancer agent containing one or more of these novelsapogenins for the treatment of cancer, particularly multi-drugresistant cancers, as well as a process for producing these novelsapogenins.

BACKGROUND OF INVENTION AND RELATED ART

[0003] Since the beginning of the last decade, anti-cancer research hasbeen increasingly directed to the discovery of novel anti-cancer agentsobtained from natural sources, as well as identifying and preparingsynthetic compounds found in natural sources.

[0004] Ginseng saponins (dammarane saponins, also called “ginsenosides”,which are effective ingredients that organically exist in panax ginseng,panax quinguefol, panax notoginseng and other species in the ginsengfamily) and sapogenins (those that do not naturally exist in the ginsengplant or other species in the ginseng family and can be derived onlythrough chemical structure modification by cleavage and/orsemi-synthesis of dammarane saponins), as natural-source root compounds,have been broadly researched for their anti-cancer characteristics. Someof them have been reported to have anti-cancer effects, of which, forexample, ginsenoside Rh2 [3-O-β-D-glucopyranosyl-20(s)-protopanaxadiol]has been reported for its anti-cancer activities [1], includinginduction of differentiation and apoptosis in cancer cells [5^(˜)11],inhibition of the growth of human ovarian cancer in nude mice after oraladministration [9], and the ability to inhibit the multiplication ofmulti-drug resistance (MDR) cancer cells while used with otherchemotherapy drugs in vitro [12].

[0005] Ginsenoside Rg3[3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-20(s)-protopanaxadiol]has been reported to inhibit the invasion by various cancer cells [13]and suppress the proliferation of human prostate cancer cells [14] invitro, and to inhibit lung metastasis in mice [15] and peritonealmetastasis in rats [16].

[0006] A metabolite of ginseng saponin produced by human intestinalbacteria, Mc [20-O-[α-L-arabinofuranosyl(1→6)-β-D-glucopyranosyl]-20(s)-protopanaxadiol], has been reported toinhibit the vascularization of tumors and extravasation of cancer cells[17].

[0007] While conventional chemotherapy agents directly attack the cancercells and exhibit severe adverse side effects, some ginseng saponins andsapogenins, as well as their intestinal bacteria metabolites, have beenreported to have inhibitory effects on cancers by induction ofcancer-cell apoptosis and/or by suppression of vascularization ofcancers with few adverse side effects.

[0008] In the case of treatment of cancers with ginseng saponins, it hasbeen reported that saponins which are metabolized to sapogenins byintestinal bacteria have anti-cancer effects. It has also been reportedthat ginseng saponins with a hydroxyl at C-20(R), or 20(R) epimers, suchas 20(R)-Rh2 and 20(R)-Rg3 have much lower biological activities thanthose with a hydroxyl at C-20(S), or 20(S) epimers, such as 20(S)-Rh2and 20(S)-Rg3 respectively. Currently, mixtures of 20(R) and 20(S)epimers are very difficult if not impossible to separate. Thus themixture has lower efficacy than that of 20(S) epimer. Furthermore, allpreviously discovered ginsenosides and sapogenins either have sugarmoieties at C-3, C-6 or C-20, or have a hydroxyl at C-20, or have both.

SUMMARY OF THE INVENTION

[0009] This invention relates to a group of novel sapogenins, their usein anti-cancer applications, and to a process for their production. Moreparticularly, this invention pertains to a novel group of dammaranesapogenins, PAM-120, PBM-110 and PBM-100 (the dammarance sapogeninestructure in these three sapogenins is specifically clean of any sugarmoieties (glycons) at any position and a hydroxyl at C-20) and PAN-20and PAN-30 (the dammarance sapogenin structure has sugar moieties(glycons) but is free of hydroxyl at C-20), obtained by chemicalcleavage of dammarane saponins. The invention also includes a novelapplication of the said sapogenins for anti-cancer treatment by usingthem separately or together, and/or jointly with other drugs, as well asto the process of producing these novel sapogenins. Said novel dammaranesapogenins show surprising anti-cancer effect when applied. Inparticular, the novel dammarane sapogenins show unexpected and superioractivity against multi-drug resistant cancers.

[0010] The invention is directed to a sapogenin according to theformula:

[0011] wherein R1 is H, glc or glc¹⁻² glc, R2 is H or OH, R3 is H or OH;and when R1, R2 and R3 are H, there are double bonds at positions 20(21)and 24(25); and when R1 is H, R2 is OH and R3 is OH, there are doublebonds at positions 20(22) and 25(26); and when R1 is H, R2 is OH and R3is H, there are double bonds at positions 20(22) and 24(25); and when R1is glc, R2 is H and R3 is H, there are double bonds at positions 20(21)and 24(25); and when RI is glc¹⁻²glc, R2 is H and R3 is H, there aredouble bonds at positions 20(22) and 24(25); and pharmaceuticallyacceptable compositions incorporating said sapogenins.

[0012] The invention in one embodiment is directed to a sapogeninaccording to the formula:

[0013] The invention in a second embodiment is directed to a sapogeninaccording to the formula:

[0014] The invention in a third embodiment is directed to a sapogeninaccording to the formula:

[0015] The invention in a fourth embodiment is directed to a sapogeninaccording to the formula:

[0016] The invention is a fifth embodiment is directed to a sapogeninaccording to the formula:

[0017] The invention also pertains to the use of a sapogenin accordingto the formula of the invention in treating cancer cells in a humanbeing suffering from cancer, comprising killing cancer cells, inducingapoptosis in cancer cells, or inhibiting multiplication of cancer cells,or any combination thereof. The sapogenins of the invention areparticularly useful in treating drug resistant cancer cells (MDR) in ahuman being suffering from cancer, comprising using the sapogeninseither singly, or in combination with one another, or in combinationwith other chemotherapy agents.

[0018] The invention also pertains to a method of treating cancer inhuman beings or other animals suffering from cancer comprisingadministering to said human beings or other animals a therapeuticallyeffective amount of a composition comprising one or more of PAM-120,PBM-100, PBM-110, PAN-20 and PAN-30.

[0019] The method can comprise a pharmaceutically effective amount ofPAM-120, PAM-100, PBM-110, PAN 20 and PAN-30, with or without one ormore pharmaceutically acceptable carriers. The active ingredient can beadministered in a dosage of between 5 micrograms to 50 grams per 1 kgbody weight per day. A preferred range is 50 micrograms to 5 grams per 1kg body weight per day. The form of the composition can be selected fromthe group consisting of an orally administrable form, an injectableform, and a topically applicable form.

[0020] The orally administrable form can be selected from the groupconsisting of a tablet, a powder, a suspension, an emulsion, a capsule,a granule, a troche, a pill, a liquid, a spirit, a syrup and a lemonade.The injectable form can be selected from the group consisting of aliquid, a suspension and a solution. The topically applicable form canbe selected from the group consisting of a drop, a paste, an ointment, aliquid, a powder, a plaster, a suppository, an aerosol, a liniment, alotion, an enema and an emulsion. The composition can be administered tohuman beings or other animals who are receiving one or more otheranti-cancer treatments. The composition can be formulated with one ormore other anti-cancer agents, for additive treatment effects, orsynergistic treatment effects on multidrug resistance cancers or anyother cancer type.

[0021] The invention also includes the incorporation of the sapogeninsaccording to the invention in foods, health foods, nutritional products,natural products and alternative medicine products.

[0022] The invention also pertains to a process of preparing a sapogeninwhich comprises producing a ginsenoside extract from plants selectedfrom the group consisting of panax ginseng, panax quinguefol and panaxnotoginseng, and proceeding according to the following steps: (a) mixingthe ginsenoside extract with water; (b) (i) mixing the ginsenosideextract and water with a short-chain (1-5 carbon) alkali-metalalcoholate solution or a hydroxide-ethanol solution to produce amixture; and (ii) placing the resultant mixture in a reaction tank sothat the resultant mixture can undergo chemical reactions under requiredhigh temperature and high pressure; or (c) (i) alternatively, mixing theginsenosides extract with ethanol; (ii) mixing the extract and ethanolwith alkali-metal alcoholates solution to produce a mixture, and (iii)placing the resultant mixture in a reaction tank so that the resultantmixture can undergo chemical reactions under required high temperatureand high pressure; (d) after the reaction is completed, collecting anintermediate product of a mix of gensenosides and sapogenins from theethanol mixture; and (e) separating the desired sapogenins from theintermediate saponin-sapogenin mixture by silica-gel-columnchromatography.

[0023] The alkali metal can be potassium or sodium. The hydroxide can besodium hydroxide or potassium hydroxide. The alkali-metal alcoholatessolution or the concentration of hydroxide-ethanol solution can be 5-50%(W/V). The alcohol can have 1-5 carbon atoms. The temperature of thereaction tank can be between 150-300° C. and the reaction pressure canbe between 2.5-8.4 MPa.

DRAWINGS

[0024] In drawings which illustrate specific embodiments of theinvention, but which should not be construed as restricting the spiritor scope of the invention in any way:

[0025]FIG. 1 illustrates a graph of tumor inhibiting effect of variousginsenosides on B16 cells.

[0026]FIG. 2 illustrates a graph of tumor inhibiting effect of variousginsenosides on drug resistant human breast cancer cells MCF7r.

[0027]FIG. 3 illustrates a plot of the synergistic effect of PAM-120with cisplatin on drug resistant human breast cancer cells MCF7r.

[0028]FIG. 4 illustrates a plot of the synergistic effect of PAM-120with taxol on drug resistant human breast cancer cells MCF7r.

[0029]FIG. 5 illustrates a graph of the therapeutic effect of PAM-120 onmouse intracranial human malignant glioma (U87) model.

[0030]FIG. 6 illustrates a graph of the therapeutic effect of PAM-120 onmouse subcutaneous human malignant glioma (U87) model.

[0031]FIG. 7 illustrates a flow chart of two processes which can be usedto obtain the sapogenins according to the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0032] Throughout the following description, specific details are setforth in order to provide a more thorough understanding of theinvention. However, the invention may be practiced without theseparticulars. In other instances, well known elements have not been shownor described in detail to avoid unnecessarily obscuring the invention.Accordingly, the specification and drawings are to be regarded in anillustrative, rather than a restrictive, sense.

[0033] This invention relates to a physically obtained group of novelcompounds as follows:

[0034] Dammara-20(21)-diene-3,12-diol (named as PAM-120);

[0035] Dammara-20(22E)-diene-3,12,24-triol (named as PBM-100);

[0036] Dammara-2-(22E)-diene-3,12,24-triol (named as PBM-110);

[0037] 3-O-β-D-glucopyranosyl-dammara-20(21)-diene-3,12-diol (named asPAN-20); and

[0038]3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-dammara-20(22E)-diene-3,12-diol(named as PAN-30).

[0039] The chemical formulas, structures and spectrum characteristics ofthe above listed novel compounds are shown on the following pages:

[0040] Sapogenin PAM-120

[0041] Dammara-20(21)-diene-3,12-diol (Named as PAM-120)

[0042] (1) Structural formula:

[0043] (2) Molecular formula: C₃₀H₅₀O₂

[0044] (3) Molecular weight: 442.723

[0045] (4) The ¹H-NMR spectrum (300 MHz, C₅D₅N) has shown signals atδ5.28 (1H, br.t), δ5.14 (1H, s), δ4.90 (1H, s), δ1.67 (3H, s), δ1.60(3H, s), δ1.23 (3H, s), δ1.06 (3H, s), δ1.03 (3H, s), δ0.95 (3H, s) andδ0.90 (3H, s).

[0046] (5) The ¹³C-NMR spectrum (75.4 MHz, C₅D₅N) has shown signals atδ39.57 (C-1), δ28.31 (C-2), δ78.02 (C-3), δ40.30 (C-4), δ56.46 (C-5),δ18.84 (C-6), δ35.46 (C-7), δ37.53 (C-8), δ51.03 (C-9), δ39.61 (C-10),δ32.76 (C-11), δ72.51 (C-12), δ48.29 (C-13), δ51.27 (C-14), δ32.68(C-15), δ27.12 (C-16), δ52.51 (C-17), δ15.91 (C-18), δ16.61 (C-19),δ155.57 (C-20), δ108.18 (C-21), δ33.91 (C-22), δ30.82 (C-23), δ125.38(C-24), δ131.24 (C-25), δ25.81 (C-26), δ17.81 (C-27), δ28.73 (C-28),δ16.34 (C-29) and δ17.06 (C-30).

[0047] Sapogenin PBM-100

[0048] Dammara-20(22E)-diene-3,12,24-triol (Named as PBM-100)

[0049] (1) Structural formula:

[0050] (2) Molecular formula: C₃₀H₅₀O₄

[0051] (3) Molecular weight: 474.721

[0052] (4) The ¹H-NMR spectrum (300 MHz, C₅D₅N) has shown signals atδ5.31 (1H, br.t), δ5.22 (1H, s), δ4.82 (1H, s), δ1.95 (3H, s), δ1.81(3H, s), δ1.66 (3H, s), δ1.64 (3H, s), δ1.47 (3H, s), δ1.19 (3H, s),δ1.06 (3H, s) and δ1.03 (3H, s).

[0053] (5) The ¹³C-NMR spectrum (75.4 MHz, C₅D₅N) has shown signals atδ39.48 (C-1), δ27.52 (C-2), δ78.48 (C-3), δ40.42 (C-4), δ61.86 (C-5),δ67.77 (C-6), δ47.69 (C-7), δ41.48 (C-8), δ50.55 (C-9), δ39.48 (C-10),δ32.02 (C-11), δ72.63 (C-12), δ50.47 (C-13), δ50.73 (C-14), δ32.69(C-15), δ27.52 (C-16), δ50.92 (C-17), δ17.80 (C-18), δ17.70 (C-19),δ140.11 (C-20), δ13.23 (C-21), δ124.63 (C-22), δ30.04 (C-23), δ78.00(C-24), δ149.90 (C-25), δ110.54 (C-26), δ17.80 (C-27), δ28.94 (C-28),δ16.56 (C-29) and δ17.14 (C-30).

[0054] Sapogenin PBM-110

[0055] Dammara-20(22E)-diene-3,6,12-triol (so named as PBM-110)

[0056] (1) Structural formula:

[0057] (2) Molecular formula: C₃₀H₅₀O₃

[0058] (3) Molecular weight: 458.722

[0059] (4) The ¹H-NMR spectrum (300 MHz, C₅D₅N) has shown signals atδ5.31(1H, br.t), δ5.51 (1H, t, J=7.2 Hz), δ2.01 (3H, s), δ1.85 (3H, s),δ1.65 (3H, s), δ1.64 (3H, s), δ1.47 (3H, s), δ1.19 (3H, s), δ1.03 (3H,s) and δ1.01 (3H, s).

[0060] (5) The ¹³ C-NMR spectrum (75.4 MHz, C₅D₅N) has shown signals atδ39.48 (C-1), δ27.52 (C-2), δ78.48 (C-3), δ40.42 (C-4), δ61.86 (C-5),δ67.77 (C-6), δ47.69 (C-7), δ41.48 (C-8), δ50.55 (C-9), δ39.48 (C-10),δ32.02 (C-11), δ72.63 (C-12), δ50.47 (C-13), δ50.73 (C-14), δ32.69(C-15), δ27.52 (C-16), δ50.92 (C-17), δ17.80 (C-18), δ17.70 (C-19),δ140. 11 (C-20), δ313.23 (C-21), δ124.63 (C-22), δ30.04 (C-23), δ124.63(C-24), δ131.33 (C-25), δ25.76 (C-26), δ17.50 (C-27), δ28.94 (C-28),δ16.56 (C-29) and δ17.14 (C-30).

[0061] Sapogenin PAN-20

[0062] 3-O-β-D-glucopyranosyl-dammara-20(21)-diene-3,12-diol (Named asPAN-20)

[0063] (1) Structural formula:

[0064] (2) Molecular formula: C₃₆H₆₀O₇

[0065] (3) Molecular weight: 604.863

[0066] (4) The ¹H-NMR spectrum (300 MHz, C₅D₅N) has shown signals atδ4.92 (1H, d, J=7.5 Hz), δ5.29 (1H, br.t), δ5.14 (1H, s), δ4.90 (1H, s),δ1.66 (3H, s), δ1.60 (3H, s), δ1.30 (3H, s), δ1.02 (3H, s), δ0.98 (3H,s), δ0.98 (3H, s) and δ0.81 (3H, s).

[0067] (5) The ¹³C-NMR spectrum (75.4 MHz, C₅D₅N) for aglycon moiety hasshown signals at δ39.34 (C-1), δ27.13 (C-2), δ88.82 (C-3), δ40.26 (C-4),δ56.47 (C-5), δ18.52 (C-6), δ35.40 (C-7), δ37.12 (C-8), δ50.91 (C-9),δ39.74 (C-10), δ32.73 (C-11), δ72.47 (C-12), δ48.30 (C-13), δ51.26(C-14), δ32.74 (C-15), δ26.78 (C-16), δ52.52 (C-17), δ15.86 (C-18),δ16.52 (C-19), δ155.58 (C-20), δ108.19 (C-21), δ33.91 (C-22), δ30.82(C-23), δ125.39 (C-24), δ131.25 (C-25), δ25.81 (C-26), δ17.81 (C-27),δ28.73 (C-28), δ16.83 (C-29) and δ17.05 (C-30). The ¹³C-NMR spectrum(75.4 MHz, C5D5N) for 3-glucopyranosyl has shown signals at δ107.00(C-1″), δ75.82 (C-2″), δ78.79 (C-3″), δ71.94 (C-4″), δ78.39 (C-5″) andδ63.14 (C-6″).

[0068] Sapogenin PAN-30

[0069]3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-dammara-20(22E)-diene-3,12-diol(Named as PAN-30)

[0070] (1) Structural formula:

[0071] (2) Molecular formula: C₄₂H₇₀O₁₂

[0072] (3) Molecular weight: 766.587

[0073] (4) The ¹³C-NMR spectrum (75.4 MHz, C5D5N) has shown signals atδ39.17 (C-1), δ28.00 (C-2), δ88.82 (C-3), δ40.14 (C-4), δ56.29 (C-5),δ18.33 (C-6), δ35.24 (C-7), δ39.60 (C-8), δ50.66 (C-9), δ36.91 (C-10),δ32.10 (C-11), δ72.49 (C-12), δ50.33 (C-13), δ50.91 (C-14), δ32.54(C-15), δ26.64 (C-16), δ50.80 (C-17), δ16.35 (C-18), δ16.49 (C-19),δ140.06 (C-20), δ13.07 (C-21), δ123.21 (C-22), δ27.35 (C-23), δ123.54(C-24), δ131.16 (C-25), δ25.60 (C-26), δ17.66 (C-27), δ28.73 (C-28),δ15.72 (C-29) and δ16.92 (C-30).

[0074] The inventors herein have discovered that the dammarancesapogenin structure that is modified to be specifically clean of anysugar moieties (glycons) at any position and free of hydroxyl at C-20has surprisingly improved effectiveness in treating cancers,particularly in treating multi-drug resistant cancers, compared tosapogenins that have sugar moieties on the structure or a hydroxyl atC-20 The inventors have unexpectedly found that PAM-120, PBM-110 andPBM-100, which all fall into this chemical category, have greateranti-cancer effect than other known saponins and sapogenins. Inparticular, these three sapogenins, and especially PAM-120, showsurprisingly effective activity in the treatment of multi-drug resistantcancers.

[0075] The inventors have also surprisingly and unexpectedly found thata dammarance sapogenin structure which is free of a hydroxyl at C-20,even though there may be a sugar moiety on the structure, demonstrateseffective anti-cancer activity, particularly in the treatment ofmulti-drug resistant cancers. PAN-20 and PAN-30, according to thisinvention, fall into this latter category.

[0076] While the inventors do not wish to be bound by any adversetheories if proved to be unfounded, the inventors offer the following asan aid in understanding the invention. It seems that sapogenins thathave no hydroxyl at C-20 compared to sapogenins that have a hydroxyl atC-20 are surprisingly effective in cancer treatment. It also seems thata sapogenin that does not have a sugar moiety (glycon) on the sapogeninstructure, is more effective than sapogenins that include a sugarmoiety. It also seems that the diol is more effective than the triol.None of this could be predicted, or forecast without testing thesapogenins of the invention.

[0077] According to this invention and varying with the severity ofsymptoms experienced by the patient, the active daily dose of sapogeninPAM-120 is 0.1 mg—10 g per kg of body weight, or preferably, 1 mg—1 gper kg of body weight. The active daily dose of sapogenin PBM-110 is 0.1mg—10 g per kg of body weight, or preferably, 1 mg—1 g per kg of bodyweight. The active daily dose of sapogenin PBM-100 is 0.1 mg—10 g per kgof body weight, or preferably, 1 mg—1 g per kg of body weight. Theactive daily dose of sapogenin PAN-20 is 0.1 mg—10 g per kg of bodyweight, or preferably, 1 mg—1 g per kg of body weight. The active dailydose of sapogenin PAN-30 is 0.1 mg—10 g per kg of body weight, orpreferably, 1 mg—1 g per kg of body weight.

[0078] The anti-cancer agent according to this invention contains one ormore of the said novel sapogenins PAM-120, PBM-100, PBM-110, PAN-20 andPAN-30, with or without other anti-cancer agent, used with or withoutone or more pharmaceutically acceptable carriers such as solid andliquid excipients.

[0079] The administration forms of the said anti-cancer agents accordingto the invention are listed as follows:

[0080] Injection forms, including but not limited to intramuscular (IM)injection, intravenous (IV) injection, subcutaneous injection andtargeted-tissue injection in aqueous solutions, oil solutions, emulsion,or any forms;

[0081] Oral forms, including but not limited to tablets, capsules,granules, pills, suspensions, powders, sprits, emulsifiers, and syrups;and

[0082] Topical form, including but not limited to drops, lotions,enemas, ointments, suspensions, paps, pastes, suppositories, aerosols,cataplasmas, emulsifiers, liniments, and plasters.

[0083] This invention also relates to a production process that can beused to commercially produce the above mentioned novel dammaranesapogenins for anti-cancer applications through chemical cleavage andsemi-synthesis of dammarane saponins.

[0084]FIG. 7 illustrates a flow sheet of two alternative processes whichcan be utilized to produce the sapogenins according to the invention.The production process according to the invention uses generalginsenosides (also called dammarane saponins including Ra, Rc, Rd, Re,etc.) extracted from plants selected from the ginseng family such aspanax ginseng, panax quinguefol and panax notoginseng as raw materials.In the process, according to this invention, general ginsenosides arefirst mixed with water and then with short-chain (1-5 carbon)alkali-metal alcoholate solution or hydroxide-ethanol solution. Themixture is then put into a reaction tank to undergo chemical reactionsunder required high temperature and high pressure. Alternatively,general ginsenosides are first mixed with ethanol, and then withalkali-metal alcoholates solution. The mixture is thereafter put into areaction tank to undergo chemical reactions under required hightemperature and high pressure. After the required period of time for thereaction to be complete, the intermediate product of a mix ofginsenosides and sapogenins are collected from the ethanol solution. Thenext step is to separate the desired dammarane sapogenins from theintermediate saponin-sapogenin mix by using silica-gel-columnchromatography. According to this invention, the said alkali metal canbe potassium or sodium, the hydroxide can be sodium hydroxide orpotassium hydroxide, the concentration of alkali-metal alcoholatessolution or the concentration of hydroxide-ethanol solution can be5^(˜)50% (W/V), and the short chain alcohol can be one with 1^(˜)5carbon atoms. In this invention, during the production process, thereaction tank's temperature can be between 150^(˜)300° C. and thereaction pressure is between 2.5^(—)8.4 MPa.

[0085] Cancers susceptible to treatment with the compounds of theinvention alone or in combination with a chemotherapeutic in accordancewith various aspects of the invention may include both primary andmetastatic tumors and hyperplasias, including carcinomas of breast,colon, rectum, lung, oropharynx, hypopharynx, esophagus, stomach,pancreas, liver, gallbladder and bile ducts, small intestine, urinarytract (including kidney, bladder and urothelium), female genital tract(including cervix, uterus, and ovaries as well as choricarcinoma andgestational trophoblastic disease)), male genital tract (includingprostate, seminal vesicles, testes and germ cell tumors), endocrineglands (including the thyroid, adrenal, and pituitary glands), and skin,as well as hemangiomas, melanomas, sarcomas (including those arisingfrom bone and soft tissues as well as Kaposi's sarcoma), and tumors ofthe brain, nerves, eyes, and meninges (including astrocytomas, gliomas,glioblastomas, retinoblastomas, neuromas, neuroblastomas, Schwannomas,and meningiomas). In some aspects of the invention, the compounds of theinvention in combination with a chemotherapeutic may also be useful intreating hematopoietic cancers such as leukemias (i.e. chloromas,plasmacytomas and the plaques and tumors of mycosis fungoides andcutaneous T-cell lymphoma/leukemia) and lymphomas (both Hodgkin's andnon-Hodgkin's lymphomas).

[0086] The compounds of the invention and the chemotherapeutic may beadministered in combination separately or as one single combinedpharmaceutical composition. The amount of each component administeredmay be determined by an attending clinician, taking into consideration avariety of factors such as the etiology and severity of the disease, thepatient's condition and age and the potency of each component. thecomponents may be administered in accordance with the standardmethodologies as, for example, disclosed in the Physician's DeskReference (PDR) published by Medical Economics Co. Inc. of Oradell, N.J.

[0087] One or more pharmaceutically acceptable carriers or exipients maybe used to formulate pharmaceutical compositions of the invention,including solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic and absorption delaying agents, and the likethat are physiologically compatible. In alternative embodiments, thecarrier may be suitable for parenteral, intravenous, intraperitoneal,intramuscular, sublingual or oral administration. Pharmaceuticallyacceptable carriers may include sterile aqueous solutions or dispersionsand sterile powders for the extemporaneous preparation of sterileinjectable solutions or dispersion. Except insofar as any conventionalmedia or agent is incompatible with the active compound, use thereof inthe pharmaceutical compositions of the invention is contemplated.Supplementary active compounds can also be incorporated into thepharmaceutical compositions.

[0088] Pharmaceutical compositions typically must be sterile and stableunder the conditions of manufacture and storage. The composition may beformulated as a solution, microemulsion, liposome, or other orderedstructure suitable to high drug concentration. The carrier can be asolvent or dispersion medium containing, for example, water, ethanol,polyol (for example, glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), and suitable mixtures thereof. The properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. In many cases, it will bepreferable to include isotonic agents, for example, sugars, polyalcoholssuch as mannitol, sorbitol, or sodium chloride in the composition.Prolonged absorption of the injectable compositions can be brought aboutthe including in the composition an agent which delays absorption, forexample, monostearate salts and gelatin. Moreover, the pharmaceuticalcompositions may be administered in a time release formulation, forexample, in a composition which includes a slow release polymer. Theactive compounds can be prepared with carriers that will protect thecompound against rapid release, such as controlled release formulation,including implants and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers(PLG). Many methods for the preparation of such formulations arepatented or generally know to those skilled in the art.

[0089] Sterile injectable solutions can be prepared by incorporating anactive compound in the required amount in an appropriate solvent withone or a combination of ingredients enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the active compound into a sterile vehicle whichcontains a basic dispersion medium and the required other ingredientsfrom those enumerated above. In the case of sterile powders for thepreparation of sterile injectable solutions, the preferred methods ofpreparation are vacuum drying and freeze-drying which yields a powder ofthe active ingredient plus any additional desired ingredient from apreviously sterile-filtered solution thereof. Pharmaceuticalcompositions may be formulated with one or more compounds that enhancethe solubility of the active compounds.

EXAMPLES Examples of Preparation Processes: Example 1 PreparationProcess of Producing PAM-120, PBM=100, and PAN-20

[0090] Ginseng crude extract 10 g was dissolved in 40 mL of 95% ethanol

[0091] Add 40 mL of 5 N NaOH

[0092] Pour into the reaction tank, and set temperature to 240C, andpressure to 3.5 Mpa, for 1.5 hours

[0093] Reduce temperature to room temperature, and take the products outthe tank

[0094] Add HCl to neutralize pH to about 7, and expend the volume to 800mL using water

[0095] Extract 3 times with acetic ester, 100 mL each time

[0096] Combine all the extracts, and reduce the pressure to dry. Thus,obtain 3.8 g of dried extracts

[0097] Grind and dissolved the extract in 20 mL of methanol, and mix themethanol solution with silica gel

[0098] Dry up the mixture, and then grind to fine powder

[0099] Load the Silica gel column

[0100] Wash the column with 60 mL of ether:petroleum benzin (1:3), andthus, 250 mg of PAM-120, and 45 mg of PBM-100 were obtained

[0101] Wash the column with 90 mL of chlorofom:methanol (95:5), and thus50 mg of PAN-20 was obtained.

Example 2 Another Example of Preparation Process ProducingPAM-120PBM-100, and PAN-20

[0102] 10 g of Ginseng crude extract was added into reaction tank

[0103] Add to the reaction tank 100 mL of 5 N NaOH

[0104] Set temperature to 270C and pressure to 4.5 Mpa for 1 hour

[0105] Reduce temperature to room temperature, then take out theproducts

[0106] Neutralize the pH to 7 using HCl

[0107] Filter and keep the solids

[0108] Dissolve the solids in 10 mL of 95% Ethanol

[0109] Add water to make ethanol content less than 5%

[0110] Sit still overnight

[0111] Filter and keep the solids

[0112] Dry the solids

[0113] Dissolved the solids in 10 mL of methanol

[0114] Filter and keep the solution

[0115] Dry the solution to obtain 3.6 g of products

[0116] Mix the products with 11 g of silica gel

[0117] Grind and then load the silica gel column

[0118] Wash the column with 100 mL of ether:petroleum benzin (1:3), andthus, 60 mg of PAM-120, and 65 mg of PBM-100 were obtained

[0119] Wash the column with chloroform:methanol (95:5), and thus 60 mgof PAN-20 was obtained.

Example 3 Comparison of Cancer Cell Inhibition Effects in vitro BetweenGinsenoside 20(S)-Rh2 and Novel Dammarane Sapogenins PAM-120, PBM-100,PAN-30 and their Composition

[0120] A. Method

[0121] Composition: 20(S)-Rh2 was provided by Shenyang PegasusPharmaceutical R&D Co., China, with a purity of over 98%. The molecularweight for Rh2 was 622.3. Sapogenins PAM-120, PBM-100 and PAN-30 werederived from the process stated in Example 1. The molecular weights ofPAM-120, PBM-100 and PAN-30 were erspectively 442.7, 474.7 and 604.9,and the purity for each of the three agents was higher than 99%. Rh2,PAM-120, PBM-100 and PAN-30 were dissolved 1 gram each separately in 100mL absolute ethanol and stored at 4° C. the agents were diluted withRPMI-1640 medium to the desired concentrations as shown in Table 1.

[0122] Cells: Human non-small-cell lung carcinoma H460 cells wereincubated in RPMI-1640 medium added with 10% fetal calf serum, 100 unitspenicillin/ml, and 100 μg streptomycin/ml in 5% CO₂ at 37° C.

[0123] In vitro treatment: H460 cells were seeded in 96-wellflat-bottomed microtest-plates at 1.2×10³ cells per well, six wells ineach group, incubated in humidified 5% CO₂ at 37° C. for 24 hours withor without the agents according to the schedule as shown below. TABLE 1DOSAGE Group Dosage (μg/ml) Control No drug — Low-dose Rh2 25 PAM-120PBM-100 PAN-30 High-dose Rh2 40 PAM-120 PBM-100 PAN-30

[0124] After the 24 hours of incubation, an equal volume of 10% formalinphosphate-buffered saline containing 0.2% crystal violet was added toeach well and left at room temperature for 20 minutes. The plates werethen washed twice with distilled water and dried at room temperature.The absorbency of the stained cells at 590 nm was then measured using anautomatic microtest-plate reader. Average absorbency of the controlwells (A_(c)) without any treatment was calculated, average absorbencyof each treatment group (A_(Ti)) was determined, and then the averagecell viability of each treatment group (V_(i)) was derived using thefollowing formula:${V_{i}\quad (\%)} = {\frac{A_{Ti}}{A_{c}} \times 100\%}$

[0125] B. Result TABLE 2 CANCER CELL VIABILITY (%) Absorbency of tTestStained Cells w/ tTest w/ Group (M ± SD) V(%) Control Rh2 Control Nodrugs .368 ± .069 100.00 Low- Rh2 .278 ± .030 78.49 P < 0.01 dose PAM-.220 ± .051 62.08 P < 0.01 P < 0.05 (25 120 μg/ PBM- .223 ± .040 62.72 P< 0.01 P < 0.05 ml) 100 PAN-30 .249 ± .045 70.30 P < 0.01 High- Rh2 .181± .049 50.99 P < 0.01 dose PAM- .125 ± .031 35.34 P < 0.01 P < 0.05 (40120 μg/ PBM- .130 ± .019 36.51 P < 0.01 P < 0.05 ml) 100 PAN-30 .147 ±.032 41.49 P < 0.01

[0126] The results in Table 2 show a significant inhibitory effect onproliferation of H460 cells by each of the novel compounds PAM-120,PBM-100 and PAN-30 (P<0.01 compared with that of the Rh2 control), and anotable increase in inhibitory effect of PAM-120 and PBM-100 on theproliferation of H460 cells (P<0.05 compared with that of Rh2).

Example 4 Tumor Weight Test

[0127] A. Method

[0128] Forty (40) C57BL/6J mice weighing 18-22 g were randomly dividedinto four groups: one control group and three treatment groups, eachwith 10 animals. Mouse sarcoma 180 cells were hyperdermicallytransplanted into the mice by using a transplantation needle under theright armpit. After the transplantation, all mice formed a tumor. Themix composition of ginsenosides and sapogenins including the three noveldammarane sapogenins (PAM-120, PBM-100 and PAN-30), derived as anintermediate product from the process described in Example 2, wasprepared into a suspension form. The mice were weighed daily prior todrug administration to determine the actual measurement of drugadministered. The drug administration started from 24 hours post tumortransplantation. The mice in the three treatment groups were orallygiven the mix composition at a daily dose of 0.4 mg/kg, 1.2 mg/kg and3.6 mg/kg respectively for 8 days using a gastric catheter. The mice inthe control group were orally given a normal saline placebo. 24 hoursafter the last administration of the drug, the mice were sacrificed withan overdose of anesthetics. The weight of the sarcoma in each mouse wasmeasured. The average tumor weight of each treatment group (Wt_(i)) andthat of the control group (Wc) were calculated, and the tumor inhibitionratio (R_(i)) of each treatment group was determined with the followingformula:${R_{i}\quad (\%)} = {\frac{{Wc} - {Wt}_{i}}{W_{c}} \times 100\%}$

[0129] B. Result TABLE 3 TUMOR WEIGHT RATIO (%) GROUP MICE# Tumor Weight(g) (M ∓ SD) R(%) P Control 10 2.995 ∓ 0.621 Mix 0.4 mg/kg 10 1.269 ∓0.525 57.63 <0.01 Mix 1.2 mg/kg 10 0.725 ∓ 0.270 75.79 <0.01 Mix 3.6mg/kg 10 0.388 ∓ 0.130 87.04 <0.01

[0130] The results in Table 3 have demonstrated that oral administrationof the subject mix composition, the intermediate product from Example 2,achieves tumor inhibition ratios of 58%, 76% and 87% respectively at thedoses of 0.4 mg/kg, 1.2 mg/kg and 3.6 mg/kg, showing a dose relatedanti-cancer efficacy of the mix of the intermediate product of Example 2containing the three novel sapogenins PAM-120, PBM-100 and PAN-30 andsome other saponins and sapogenins whose structures are known orunknown.

Example 5 Cancer Bearing Mice Life Prolongation Test

[0131] A. Method

[0132] Fifty (50) C57BL/6J mice weighing 18-22 g, without sexdiscrimination, were randomly assigned to one control group and fourtreatment groups, each with 10 animals. Murine sarcoma 180 cells wereintraperitoneally transplanted to the mice. 20(S)-Rh2 was provided byShenyang Pegasus Pharmaceutical R&D Co., China, with a purity of over98%. The molecular weight for Rh2 was 622.3. The novel sapogenin PAM-120was derived from the process described in Example 2 according to thisinvention. The molecular weight of PAM-120 was 442.7, and the purity forPAM-120 was higher than 99%. The drugs were prepared into a suspensionform respectively. The mice were weighed daily prior to drugadministration to determine the actual measurement of drug administered.Drug administration started from 24 hours post tumor inoculation. Themice in the two low-dose treatment groups were orally given the Rh2 andPAM-120 preparations using a gastric catheter at a daily dose of 10mg/kg of Rh2 and 10 mg/kg of PAM-120 respectively for a lifetime or upto 120 days. The mice in the two high-dose treatment groups were orallygiven the Rh2 and PAM-120 preparations using a gastric catheter at adaily dose of 25 mg/kg of Rh2 and 25 mg/kg of PAM-120 respectively for alifetime or up to 120 days. The mice in the control group were orallygiven a normal saline. For each group, the days of survival for 50%animals (DS₅₀) and the average days of survival (ADS) were recorded. Forgroups containing one or more animals that could have lived longer than120 days (the designed sacrifice day was d 120), the ADS would be socalculated that these animals were counted as if they had died on day120, and a note would be made. The life prolongation rate (LPR) wascalculated with the following formula:${{LPR}\quad (\%)} = {\frac{{ADS}_{({treatment})} - {ADS}_{({control})}}{{ADS}_{({control})}} \times 100\%}$

[0133] B. Result TABLE 4 B16 MELANOMA BEARING MICE LIFE PROLONGATIONRATE % Group DS₅₀ ADS(M ∓ SD) LPR(%) Control 14 14.7 ∓ 5.4  Rh2 (3mg/kg) 22 24.7 ∓ 12.6 68.0 PAM-120 (3 mg/kg) 38 38.6 ∓ 16.4 162.6 Rh2 (6mg/kg) 41 44.3 ∓ 19.6 201.4 PAM-120 (6 mg/kg) 77 80.6 ∓ 34.4 448.3

[0134] The anti-cancer effect of the novel sapogenin PAM-120 wasindicated by the significant increase in life prolongation of the micebearing murine sarcoma (P<0.01 compared with the average days ofsurvival in the control). Better anti-cancer effect of novel sapogeninPAM-120 on murine sarcoma than that of Rh2 was demonstraated by thesignificant increase in life prolongation of the sarcoma-bearing mice(P<0.01, compared with the average days of survival in the relevant Rh2treatment dose groups). Two mice in the 25 mg/kg composition treatmentgroup survived for the whole 120 days, and were found to have no tumorswhatsoever existing in their bodies postmortally.

[0135]FIG. 1 illustrates a graph of tumor inhibiting effect of variousginsenosides on B16 cells. Mouse melanoma tumor B16 cells were culturedwith DMEM and 5% serum supplement in 96-well dishes. Cells were thentreated with various concentrations of PAN-20, PAN-30, PBM-100, PBM-110,PAM-120 and Rh2, respectively. The number of alive cells were measuredusing MTT method 24 hours after the treatment and compared with thecontrol samples. All the new compounds showed a significantly highertumor inhibitory effect than RH2, especially at low concentrations(p<0.01, Student t test).

[0136]FIG. 2 illustrates a graph of tumor inhibiting effect of variousginsenosides on drug resistant human breast cancer cells MCF7r. Humandrug resistant breast cancer cells (MCF7r) were cultured with DMEM and5% serum supplement in 96-well dishes. Cells were then treated withvarious concentrations of PAN-20, PAN-10, PAN-12 and Hr2, respectively.The number of alive cells were measured using MTT method 24 hours afterthe treatment and compared with the control samples. All new compoundsshowed a significantly higher tumor inhibitory effect than Rh2,especially at low concentrations (p<0.01, Student t test).

[0137]FIG. 3 illustrates a plot of the synergistic effect of PAM-120with Cisplatin on drug resistant human breast cancer cells MCF7r. MCF7rcells were treated with anti-cancer chemotherapy agent Cisplatin atvarious concentrations in the presence of 10 ug/ml PAM-120. In FIG. 3,the first bars in each concentration group represent percentages ofalive cells 24 hours after treatment with Cisplatin only. The secondbars in each group represent the results of cells treated with Cisplatinand PAM-120.

[0138]FIG. 4 illustrates a plot of the synergistic effect of PAM-120with Taxol on drug resistant human breast cancer cells MCF7r. MCF7rcells were treated with anti-cancer chemotherapy agent Taxol at variousconcentrations in the presence of 10 ug/ml PAM-120 or 20 ug/ml RH2. InFIG. 4, the first bars in each concentration group represent percentagesof alive cells 24 hours after treatment with Taxol only. The second barsin each group represent the results of cells treated with Taxol andPAM-120, while the third bars represent the results of cells treatedwith 20 ug/ml Rh2.

[0139]FIG. 5 illustrates a graph of the therapeutic effect of PAM-120 onmouse intracranial human malignant glioma (U87) model. Nude mice wereintracranially implanted with human malignant glioma cells (U87). On day10 post tumor implantation, animals were treated with various dosages ofPAM-120. Animals treated with 25 mg/kg and 50 mg/kg PAM-120 hadsignificantly longer survival time after tumor implantation (p<0.01,Kaplan Meier analysis).

[0140]FIG. 6 illustrates a graph of the therapeutic effect of PAM-120 onmouse subcutaneous human malignant glioma (U87) model. Nude mice weresubcutaneously implanted with human malignant glioma cells (U87). On day7 post tumor implantation, animals were treated with 25 mg/ml PAM-120 orequal dose of Rh2. Tumor sizes were measured on day 7 (before treatment)and day 24 (after the treatment). Both PAM-120 and Rh2 significantlyinhibited tumor growth comparing to PBS control animals. Tumor sizes inPAM-120 treated animals were significantly smaller than those in the Rh2treated animals (p<0.05).

[0141]FIG. 7 illustrates a flow chart of two processes which can be usedto obtain the sapogenins according to the invention.

[0142] As will be apparent to those skilled in the art in the light ofthe foregoing disclosure, many alterations and modifications arepossible in the practice of this invention without departing from thespirit or scope thereof. Accordingly, the scope of the invention is tobe construed in accordance with the substance defined by the followingclaims.

CITED REFERENCES

[0143] 1. Kim N D, Park M K, Lee S K, Park J H, Kim J M (1998) Processedginseng product with enhanced pharmacological effects. U.S. Pat. No.5,776,860.

[0144] 2. Lee Y N, Lee H Y, Chung H Y, Kim S I, Lee S K, Park B C, Kim KW (1996) In vitro induction of differentiation by ginsenosides in F9teratocarcinoma cells. Eur J Cancer. 1820-8

[0145] 3. Odashima S, Ohta T, Kohno H, Matsuda T, Kitagawa I, Abe H,Arichi S (1985) Control of phenotypic expression of cultured B16melanoma cells by plant glycosides. Cancer Res. 85: 2781-8

[0146] 4. Xia L J, Han R (1996) [Differentiation of B16 melanoma cellsinduced by ginsenoside RH2]. Yao Hsueh Hsueh Pao. 31: 782-5

[0147] 5. Kikuchi Y, Sasa H, Kita T, Hirata J, Tode T, Nagata I (1991)Inhibition of human ovarian cancer cell proliferation in vitro byginsenoside Rh2 and adjuvant effects to cisplatin in vivo. Anti-cancerDrugs. 2: 63-7

[0148] 6. Lee K Y, Park J A, Chung E, Lee Y H, Kim S I, Lee S K (1996)Ginsenoside-Rh2 blocks the cell cycle of SK-HEP-1 cells at the G1/Sboundary by selectively inducing the protein expression of p27kip1.Cancer Lett. 110: 193-200

[0149] 7. Oh M, Choi Y H, Choi S, Chung H, Kim K, Kim S I, Kim D K, KimN D (1999) Anti-proliferating effects of ginsenoside Rh2 on MCF-7 humanbreast cancer cells. Int J Oncol. 18: 869-75

[0150] 8. Ota T, Maeda M, Odashima S, Ninomiya T J, Tatsuka M (1997) G1phase-specific suppression of the Cdk2 activity by ginsenoside Rh2 incultured murine cells. Life Sci. 60: PL39-88

[0151] 9. Nakata H, Kikuchi Y, Tode T, Hirata J, Kita T, Ishii K, KudohK, Nagata I, Shinomiya N (1998) Inhibitory effects of ginsenoside Rh2 ontumor growth in nude mice bearing human ovarian cancer cells. Jpn JCancer Res. 89: 733-80

[0152] 10. Kim H E, Oh J H, Lee S K, Oh Y J (1999) Ginsenoside RH-2induces apoptotic cell death in rat C6 glioma via a reactive oxygen- andcaspase-dependent but Bcl-X(L)-independent pathway. Life Sci. 65:PL33-80

[0153] 11. Park J A, Lee K Y, Oh Y J, Kim K W, Lee S K (1997) Activationof caspase-3 protease via a Bcl-2-insensitive pathway during the processof ginsenoside Rh2-induced apoptosis. Cancer Lett. 121: 73-81

[0154] 12. Jia W (2000) Ginsenoside Chemotherapy. U.S. PatentProvisional File—Serial No. 60/204/765

[0155] 13. Shinkai K, Akedo H, Mukai M, Imamura F, Isoai A, Kobayashi M,Kitagawa I (1996) Inhibition of in vitro tumor cell invasion byginsenoside Rg3. Jpn J Cancer Res. 87: 357-62.

[0156] 14. Liu W K, Xu S X, Che C T (2000) Anti-proliferative effect ofginseng saponins on human prostate cancer cell line. Life Sci.67(11):1297-306.

[0157] 15. Iishi H, Tatsuta M, Baba M, Uehara H, Nakaizumi A, Shinkai K,Akedo H, Funai H, Ishiguro S, Kitagawa I (1997) Inhibition byginsenoside Rg3 of bombesin-enhanced peritoneal metastasis of intestinaladenocarcinomas induced by azoxymethane in Wistar rats. Clin ExpMetastasis 15: 603-11.

[0158] 16 Mochizuki M, Yoo Y C, Matsuzawa K, Sato K, Saiki I, Tono-okaS, Samukawa K, Azuma I (1995) Inhibitory effect of tumor metastasis inmice by saponins, ginsenoside-Rb2, 20(R)- and 20(S)-ginsenoside-Rg3, ofred ginseng. Biol Pharm Bull. 18: 1197-202.

[0159] 17. Hasegawa H, Jong H S, Matsumiya S, Uchiyama M, Jae D H (1999)Metabolites of Ginseng Saponins by Human Intestinal Bacteria and ItsPreparation for an Anti-cancer. U.S. Pat. No. 5,919,770.

What is claimed is:
 1. A sapogenin according to the formula:

wherein R1 is H, glc or glc¹⁻² glc, R2 is H or OH, R3 is H or OH; andwhen R1, R2 and R3 are H, there are double bonds at positions 20(21) and24(25); and when R1 is H, R2 is OH and R3 is OH, there are double bondsat positions 20(22) and 25(26); and when R1 is H, R2 is OH and R3 is H,there are double bonds at positions 20(22) and 24(25); and when R1 isglc, R2 is H and R3 is H, there are double bonds at positions 20(21) and24(25); and when R1 is glc¹⁻²glc, R2 is H and R3 is H, there are doublebonds at positions 20(22) and 24(25); and pharmaceutically acceptablecompositions incorporating said sapogenins.
 2. A sapogenin as claimed inclaim 1 wherein R1, R2 and R3 are H, and there are double bonds at20(21) and 24(25).
 3. A sapogenin as claimed in claim 1 wherein R1 is H,R2 and R3 are OH, and there are double bonds at 20(22) and 25(26).
 4. Asapogenin as claimed in claim 1 wherein R1 is H, R2 is OH and R3 is H,and there are double bonds at 20(22) and 24(25).
 5. A sapogenin asclaimed in claim 1 wherein R1 is glc, R2 and R3 are H, and there aredouble bonds at 20(21) and 24(25).
 6. A sapogenin as claimed in claim 1wherein R1 is glc¹⁻²glc, R2 and R3 are H, and there are double bonds at20(22) and 24(25).
 7. The use of a sapogenin according to the formularecited in claim 1 in treating cancer cells in a human being sufferingfrom cancer, comprising killing cancer cells, inducing apoptosis incancer cells, or inhibiting multiplication of cancer cells, or anycombination thereof.
 8. The use of a sapogenin according to the formularecited in claim 1 in treating multi-drug resistant cancer cells (MDR)in a human being suffering from cancer, comprising using the sapogeninseither singly, or in combination with one another, or in combinationwith other chemotherapy agents.
 9. A sapogenin according to the formula:


10. A sapogenin according to the formula:


11. A sapogenin according to the formula:


12. A sapogenin according to the formula:


13. A sapogenin according to the formula:


14. A method of treating cancer in human beings or other animalssuffering from cancer comprising administering to said human beings atherapeutically effective amount of a composition comprising one or moreof PAM-120, PBM-100 and PBM-110.
 15. A method of treating cancer inhuman beings or other animals suffering from cancer comprisingadministering to said human beings a therapeutically effective amount ofa composition comprising one or more of PAN-20 and PAN-30.
 16. Thecancer-treatment method of claim 14 comprising a pharmaceuticallyeffective amount of PAM-120, PAM-100 and PBM-110 with or without one ormore pharmaceutically acceptable carriers, and one or morechemotherapeutic agents.
 17. The cancer-treatment method of claim 14,wherein the active ingredient is administered in a dosage of between 5micrograms to 50 grams per kg body weight per day.
 18. Thecancer-treatment method of claim 14, wherein the active ingredient isadministered in a dosage of between 50 micrograms to 5 grams per kg bodyweight per day.
 19. The cancer-treatment method of claim 17, wherein theform of the composition is selected from the group consisting of anorally administrable form, an injectable form, and a topicallyapplicable form.
 20. The cancer-treatment method of claim 19, whereinthe orally administrable form is selected from the group consisting of atablet, a powder, a suspension, an emulsion, a capsule, a granule, atroche, a pill, a liquid, a spirit, a syrup and a lemonade.
 21. Thecancer-treatment method of claim 19, wherein the injectable form isselected from the group consisting of a liquid, a suspension and asolution.
 22. The cancer-treatment method of claim 19, wherein thetopically applicable form is selected from the group consisting of adrop, a paste, an ointment, a liquid, a powder, a plaster, asuppository, an aerosol, a liniment, a lotion, an enema and an emulsion.23. The cancer-treatment method of claim 14, wherein the composition isadministered to human beings who are receiving one or more otheranti-cancer treatments.
 24. The cancer-treatment method of claim 15,wherein the composition is administered to human beings who arereceiving one or more other anti-cancer treatments.
 25. Thecancer-treatment method in claim 14, wherein the composition isformulated with one or more other anti-cancer agents, for additivetreatment effects, or synergistic treatment effects on multi-drugresistance cancers or any other cancer type.
 26. The cancer-treatmentmethod in claim 15, wherein the composition is formulated with one ormore other anti-cancer agents, for additive treatment effects, orsynergistic treatment effects on multi-drug resistance cancers or anyother cancer type.
 27. A process of preparing a sapogenin as claimed inclaim 1 which comprises producing a ginsenoside extract from plantsselected from the group consisting of panax ginseng, panax quinguefoland panax notoginseng, or a sapogenin source from some other plant, andproceeding according to the following steps: (a) mixing the ginsenosideextract with water; (b) (i) mixing the ginsenoside extract and waterwith a short-chain (1-5 carbon) alkali-metal alcoholate solution or ahydroxide-ethanol solution to produce a mixture; and (ii) placing theresultant mixture in a reaction tank so that the resultant mixture canundergo chemical reactions under required high temperature and highpressure; or (c) (i) alternatively, mixing the ginsenosides extract withethanol; (ii) mixing the extract and ethanol with alkali-metalalcoholates solution to produce a mixture, and (iii) placing theresultant mixture in a reaction tank so that the resultant mixture canundergo chemical reactions under required high temperature and highpressure; (d) after the reaction is completed, collecting anintermediate product of a mix of gensenosides and sapogenins from theethanol mixture; and (e) separating the desired sapogenins from theintermediate saponin-sapogenin mixture by silica-gel-columnchromatography.
 28. A process as claimed in claim 27 wherein the alkalimetal can be potassium or sodium.
 29. A process as claimed in claim 27wherein the hydroxide can be sodium hydroxide or potassium hydroxide.30. A process as claimed in claim 27 wherein the alkali-metalalcoholates solution or the concentration of hydroxide-ethanol solutionis 5-50% (W/V).
 31. A process as claimed in claim 27 wherein the ethanolhas 1-5 carbon atoms.
 32. The process as claimed in claim 27 wherein thetemperature of the reaction tank is between 150-300° C. and the reactionpressure is between 2.5-8.4 MPa.
 33. A process of preparing a sapogeninas claimed in claim 1 which comprises producing a ginsenoside extractfrom plants selected from the group consisting of panax ginseng, panaxquinguefol and panax notoginseng, and proceeding according to thefollowing steps: (a) mixing the ginsenoside extract with water; (b)mixing the ginsenoside extract and water with a short-chain (1-5 carbon)alkali-metal alcoholate solution or a hydroxide-ethanol solution toproduce a mixture; and (c) placing the resultant mixture in a reactiontank so that the resultant mixture can undergo chemical reactions underrequired high temperature and high pressure; or (d) after the reactionis completed, collecting an intermediate product of a mix ofgensenosides and sapogenins from the ethanol mixture; and (e) separatingthe desired sapogenins from the intermediate saponin-sapogenin mixtureby silica-gel-column chromatography.
 34. A process of preparing asapogenin as claimed in claim 1 which comprises producing a ginsenosideextract from plants selected from the group consisting of panax ginseng,panax quinguefol and panax notoginseng, and proceeding according to thefollowing steps: (a) mixing the ginsenoside extract with water; (b)alternatively, mixing the ginsenosides extract with ethanol; (c) mixingthe extract and ethanol with alkali-metal alcoholates solution toproduce a mixture, and (d) placing the resultant mixture in a reactiontank so that the resultant mixture can undergo chemical reactions underrequired high temperature and high pressure; (e) after the reaction iscompleted, collecting an intermediate product of a mix of gensenosidesand sapogenins from the ethanol mixture; and (f) separating the desiredsapogenins from the intermediate saponin-sapogenin mixture bysilica-gel-column chromatography.